粉防己碱上调PTEN表达逆转肝细胞癌细胞顺铂耐药的研究

Study of tetrandrine upregulating PTEN expression to reverse cisplatin resistance in hepatocellular carcinoma cells

目的探究粉防己碱上调人第10号染色体缺失的磷酸酶(PTEN)表达对逆转肝细胞癌细胞顺铂耐药的作用。方法将Huh7/DDP细胞分为空白对照组、NC组、a组、b组。空白对照组不接受任何处理,仅用正常培养基培养;NC组在正常培养基中加入16μmol·L^(-1)顺铂;a组在正常培养基中加入16μmol·L^(-1)顺铂,添加5μmol·L^(-1)粉防己碱;b组在正常培养基中加入16μmol·L^(-1)顺铂,并添加5μmol·L^(-1)粉防己碱,同时过表达PTEN。用细胞计数试剂盒-8(CCK-8)法评估细胞存活率,用免疫荧光法检测PTEN基因的表达情况,以及多药耐药相关蛋白(MRP)-3和MRP-5的表达情况。结果空白对照组、NC组、a组和b组的细胞存活率分别为(98.74±9.26)%、(55.08±6.38)%、(35.17±4.87)%和(24.06±4.16)%,PTEN表达水平分别为1.00±0.13、1.03±0.13、1.58±0.17和2.18±0.24,MRP-3蛋白表达水平分别为1.00±0.08、1.03±0.14、0.71±0.07和0.48±0.04,MRP-5蛋白表达水平分别为1.00±0.11、0.97±0.12、0.63±0.07和0.40±0.05。空白对照组的上述指标与NC组比较,a组的上述指标与b组比较,在统计学上差异均有统计学意义(均P<0.05)。结论粉防己碱通过上调PTEN表达来逆转肝细胞癌细胞顺铂耐药的作用,并发现其能抑制耐药相关蛋白MRP-3和MRP-5的表达。

Objective To investigate the effect of tetrandrine on reversing cisplatin resistance in hepatocellular carcinoma cells by up-regulating phosphatase and tensin homolog deleted on chromosome ten(PTEN)expression.Methods Huh7/DDP cells were divided into control group,NC group,group a and group b.The control group received no treatment and was cultured with normal medium only.In NC group,16μmol·L^(-1)cisplatin was added into normal medium.In group a,16μmol·L^(-1)cisplatin and 5μmol·L^(-1)tetrandrine were added into normal medium.In group b,16μmol·L^(-1)cisplatin and 5μmol·L^(-1)tetrandrine were added to normal medium,and PTEN was overexpressed.Cell survival was assessed by cell counting kit-8(CCK-8)method,and PTEN gene expression was detected by immunofluorescence.The impact on multidrug resistance associated protein(MRP)-3 and MRP-5 were observed.Results The cell survival rates of control group,NC group,a group and b group were(98.74±9.26)%,(55.08±6.38)%(35.17±4.87)%and(24.06±4.16)%;the expression of PTEN gene were 1.00±0.13,1.03±0.13,1.58±0.17and 2.18±0.24;the expression of MRP-3 protein were 1.00±0.08,1.03±0.14,0.71±0.07 and 0.48±0.04;MRP-5 protein expression were 1.00±0.11,0.97±0.12,0.63±0.07 and 0.40±0.05,respectively.Comparedbetween NC group and control group,compared between group a and group b,there were statistically significantdifferences in the above indexes(allP<0.05).Conclusion Tetrandrine reverses cisplatin resistance inhepatocellular carcinoma cells by up-regulating PTEN expression and inhibits the expression of drug resistance-related proteins MRP-3 and MRP-5.